Giancarlo - put these in reverse chronological order. - Dr. B
Intoduction to PCR Protocol
Week 1: First Day
1) Prepare Template Dilutions for PCR
2) Run Dilutions though PCR machine (overnight).
Week 1: Second Day
3) Create Agarose Gel
4) Run Gel for 40 minutes
Figure 1:
PCR-1 Gel Run: This gel run was done incorrectly due to the
fact that only two lanes are visible. One needs five lanes
visible for it to be done correctly.
Week 2: (PCR 1 didn't work)
-First Day
-Enzyme Digest
-Redid PCR-1
-prepare template dilutions for PCR
-run dilutions through PCR machine.
-Second Day
-PCR-1
-create Agarose gel
-run PCR dilutions through PCR gel for 40 minutes
-take picture of finished gel
Figure 2:
Week 3:
Day1:
-PCR-2
-create plasmid dilutions
-create mater mix
-create templates
-add TAQ polymerase to each template
-run templates through PCR machine
-Enzyme Digest Gel Run
Figure 3: (Enzyme Digest)
Day 2:
-run PCR-2 templates through gel
-take picture
Figure 4: (PCR2) (Failed)
This gel sample failed because only the DNA ladder appeared.
If done correctly, the DNA ladder and the rest of the 4 samples
would have appeared.
Week 4:
Day 1:
-attempted to do VirtualRefresher, but computer froze and runs won't run.
Day 2:
-Since PCR 2 failed the first time, I had to do it again.
-Run samples through PCR machine
-Make gel
-Run samples through gel
Figure 5: (PCR2)(Failed AGAIN!)
Week 6:
Day 1:
-started primer overlap
-attempted to do Virtual Refresher, but only the 4th and 5th folder didn't run (check again tomorrow)
Week 7: (during week 6 I attended lab for only 6 hours, so I'm continuing Virtual Refresher and Primer overlap)
-Secondary PCR for Primer Overlap
-Gel Run for Primary and Secondary PCR's
-I ran the 4th and 5th folders for Virtual Refresher
-Gel run for Primary and Secondary PCR (Figure below: FAILED)
Week 8:
-Since Primary and Secondary PCR gel run FAILED, I had to redo the whole primer overlap lab.
Giancarlo's Research
Giancarlo - put these in reverse chronological order. - Dr. B
Intoduction to PCR Protocol
Week 1: First Day
1) Prepare Template Dilutions for PCR
2) Run Dilutions though PCR machine (overnight).
Week 1: Second Day
3) Create Agarose Gel
4) Run Gel for 40 minutes
Figure 1:
PCR-1 Gel Run: This gel run was done incorrectly due to the
fact that only two lanes are visible. One needs five lanes
visible for it to be done correctly.
Week 2: (PCR 1 didn't work)
-First Day
-Enzyme Digest
-Redid PCR-1
-prepare template dilutions for PCR
-run dilutions through PCR machine.
-Second Day
-PCR-1
-create Agarose gel
-run PCR dilutions through PCR gel for 40 minutes
-take picture of finished gel
Figure 2:
Week 3:
Day1:
-PCR-2
-create plasmid dilutions
-create mater mix
-create templates
-add TAQ polymerase to each template
-run templates through PCR machine
-Enzyme Digest Gel Run
Figure 3: (Enzyme Digest)
Day 2:
-run PCR-2 templates through gel
-take picture
Figure 4: (PCR2) (Failed)
This gel sample failed because only the DNA ladder appeared.
If done correctly, the DNA ladder and the rest of the 4 samples
would have appeared.
Week 4:
Day 1:
-attempted to do VirtualRefresher, but computer froze and runs won't run.
Day 2:
-Since PCR 2 failed the first time, I had to do it again.
-Run samples through PCR machine
-Make gel
-Run samples through gel
Figure 5: (PCR2)(Failed AGAIN!)
Week 6:
Day 1:
-started primer overlap
-attempted to do Virtual Refresher, but only the 4th and 5th folder didn't run (check again tomorrow)
Week 7: (during week 6 I attended lab for only 6 hours, so I'm continuing Virtual Refresher and Primer overlap)
-Secondary PCR for Primer Overlap
-Gel Run for Primary and Secondary PCR's
-I ran the 4th and 5th folders for Virtual Refresher
-Gel run for Primary and Secondary PCR (Figure below: FAILED)
Week 8:
-Since Primary and Secondary PCR gel run FAILED, I had to redo the whole primer overlap lab.